OBJECTIVE: We evaluated the effect of different concentrations of the esterase inhibitor, AEBSF, and acid treatment on acyl-ghrelin stability in human plasma samples subjected to a freeze/thaw cycle.
MATERIALS AND METHODS: Four plasma samples were collected from each donor and treated with the following concentrations of AEBSF: 2 mg/ml, 1 mg/ml, 0.6 mg/ml, and 0 mg/ml. For each plasma tube collected, half of the aliquots were treated with HCl and stored at -80°C before measuring acyl-ghrelin concentration using enzyme-linked immunosorbent assay (ELISA).
RESULTS: Treatment with 1 mg/ml AEBSF + HCl resulted in significantly higher acyl-ghrelin levels compared to all other treatments except 2 mg/ml AEBSF + HCl or 0.6 mg/ml AEBSF + HCl. While all HCl-treated samples had higher acyl-ghrelin levels than their AEBSF-matched un-acidified samples, only samples treated with 1 mg/ml AEBSF significantly differed in acyl-ghrelin levels as a result of HCl treatment.
CONCLUSIONS: Our results suggest the use of 1 mg/ml AEBSF with HCl for optimal acyl-ghrelin stability in human plasma samples subjected to a freeze/thaw cycle before assay. Given that 2 mg/ml and 0.6 mg/ml AEBSF + HCl did not significantly differ from 1 mg/ml AEBSF + HCl, our data suggest that the use of AEBSF with HCl more potently prevents de-acylation of ghrelin than either treatment alone.Free PDF Download
To cite this article
S.L. Deschaine, L. Leggio
Understanding plasma treatment effect on human acyl-ghrelin concentrations
Eur Rev Med Pharmacol Sci
Vol. 24 - N. 3